Process of breeding microbes.



0. SCHMIDT.

PROGESS 0P BREEDING MIGROBES.

APPLICATION FILED MAR. 25, 1905.

94 a Patented June 6, 1911.

the

OTTO $CHMIID'1, 0F COLOGNE, GERMANY.

PROCESS OF BREEDING MICROIBES.

' Specification of Letters Patent.

Patented Juned, 1911.

application filed March 25, 1905. Serial No. 252,072.

To all whom it may concern:

Be it known that I, OTTO SCHMIDT, a subject of the German Emperor, and resident of Cologne, Ggrmany, have invented certain new and useful Improvements in Processes of Breeding Microbes, of which the following is a specification.

For the immunization of men or animals against disease-germs small quantities of pure breeds of those parasites (bacteria and other microbes) against which men or animals are to be rotected are required. Much larger quantities are required if it is intended to produce in the animal body antitoxic or bactericidal serum and to actively inoculate or immunize animals in such as high degree that their serum can be used for passive immunization of other organisms. y

The breeding of pure specimens of the bacteria, and so on, takes place on an artificially produced culture medium, the constituents of which are empirically so selected Y as experience has shown them to be the best till for the respective kind. For while a large number 01 them grow on or in all culture mediums which contain nitrogen and carbon in suitable combinations, others are far more particular in their choice, and among these especially the pathogenic kinds, which are of importance for the purpose of immunization; some of them even only grow on culture mediums of quite a definite composition. Whether the 4 culture medium is solid or liquid matters on the whole very little; practical wants generally decide this point. Besides the nourishment an important role is played also by air, temperature and light in all processes of breeding bacteria or parasitically livin micro-organisms, as some only grow if a1r and light are admitted, and others it air and light are excluded, and whereas the majority grow best at a temperature equal to that of the body, a minority grow better at a lower temperature; but with all breeding processes the main thing remains the kind and composition of the culture medium. Each pure cultivation is nothing else but the saprophytic stage of parasitic micro-organisms, as tar as under pure cultivation the breeding of parasitic micro -organisms on a dead organic substance is understood. The inanimate culture medium can never ofi'er the same conditions of existence as the living'organisms; to this is to be ascribed the fact that the germs of a be used for the production of a curative serum. I

The object of the present process is: 1, in general to breed already known but hitherto not cultivated and also hitherto unknown parasitic disease-germs belonging to the obligate parasites; 2, to breed and maintain in a pure state the so-called tacultative parasites in-undiminished virulence outside of the body. This process is based on the consideration that these breedings can only be successful outside of the human or animal body if the respective parasites can, also when bred artificially, develop purely parasitically in the tissue of a living organism, and it consists in odering to the parasites, instead of the living animals, living plants as a culture medium. The plants best suited for this are, for obvious reasons, not the Phamrogame'a but the Cryptoga-mz'a, and of these in the first place the lowest kinds living in the water, Whose soft spores ofi'er no resistance to the, entry of the parasites, but whose plasma oiier sufiicient nourishment to the parasites to thrive in it. .Which plant is the most suitable in every particular case for the respective parasite only experience and experiments can show.

The process can be carried out as follows: in order to breed obligate parasites small and well protected pieces of the diseased tissue are, before other micro orgamsms en ter, placed directly into the llquid in which the plant is in lively sporulation, or a so called tissue-culture is first made in the 111- cubator and this is,after some time, when the parasite has multiplied inthe tissue, eventually after the gradual adaptation ot the tissue to the room-temperature necessary for the plant-to be added to the culture medium of the plant. With the facultative parasites the same process may be applied, but the saprophytic growth can first repeated until the culture medium is exhausted and the plant dies. At this stage the product of the culture is ready for use, and, after passing through a coarse filter for the purpose of eliminating the more voluminous particles of the plant, it may be used for injections. The particles of the plant still mixed with the parasites may without hesitation be injected with them, for it has been found that they are rapidly absorbed without doing any harm.

By this process, 1 have succeeded in breeding in a plant the germ of an important group of diseases, the malignant tumors lupus, cankers and the like, which .I have proved to be caused by a-parasite belonging to the Protozoa. I have given a description of said parasite and a statement of my experiences and successes, together with a number of drawings and .photo graphs in my book: Mittez'lungen aus D1. bchmz'dts Laboratom'um fdr Krebsforscinmg, Bd. 1 especially in its first chapter and titled bber das Voricom/men eines protozoonartigen Parasiten in den malignen T-umoren und seine Kultw" ausserkalb des Tierlab'rpers. This animal is to be considered as being an amoeba, owing to its appearance, the manner of its motion and its propagation. If not colored, it is, in the state of rest, a round, in the state of motion,

.- an irregular shaped, limpid structure. For

of the semi-fluid body of the animal.

its motion limblike processes or pseudopodia are protruded, which are not organic, but form only processes or protuberances In sporulating sporocysts' are formed, the contents of which break up into a number of little bodies. Said parasite, which I have named by myself Illerz'stemamaligna brought upon the usual culture mediums, generally upon a dead culture medium, perishes immediately. But it develops and grows in the spores and fruits of a certain moldfungus, named Mucor racemosus, to which the said parasite is added in the above described manner. Also other fungi are able to serve as culture-plants for the said parasite. Said fungi, and specially the named mold-fungus, Vegetate in about all culture mediums today in use for breeding microbes. As a culture medium most suitable for that purpose I have found the so-called peptone-sugar-bouillon or broth.

The annexed drawing shows in Figure 1 the mold 2112mm" mcemosus in. an enlarged scale, and in Fig. 2 the parasite vegetatlng in and with the mold spores in different conditions and forms, this representation being taken from photographs.

1 shows a swarmer (1 contained in-the Mucor spore b and growing so as to form an amoeba, the protoplasm of the ucor spore being completely liquefied; 2 shows an amoeba 0 encapsuled, completely filling out,

the Mucor spore; the cyst contains, besides the rest body d, two great and some little germs e; 3 shows the same, as figure 2, but 7 the germs and rest body being upon the point to leave the double casing; 4 shows an amoeba f growing in the plasma 9 of 1 k lying tightly to to secure by United States Letters Patent 1. The hereinbefore described 'proce'ss of breeding in pure state microbes which vegetate as parasites in the human and animal body, outside of the animal body and in large quantities suflicient forthe immuniza- .131011 of other organisms, consisting in the following steps: the microbes are brought upon and into living plants of the lowestkind, which plants are brought into a suitable culture medium and made to vegetate therein, so that the microbes are made to enter the spores of the plant and to vegetate and multiply therein repeatedly, until the plant dies.

2. A special kind of means for the immunization of men against the malignant tumors, consisting in a filtered liquid containing admixed the pure breeds of those parasites which cause the said diseases, together with the spores and fruits of a certain mold-fungus, named Mucor racem'osus upon which the parasites were bred, substantially as and for the purpose set forth.

In testimony whereof I have signed my name to this specification in the presence of two subscribing witnesses.

V OTTO SCHMIDT.

Signed in presence of WILLIAM KU'EPPERS,

CHARLES LE SIMPLE. 

